*Target (protein/gene name): Eonyl -(acyl carrier protein) reductase *NCBI Gene # or RefSeq#:208435803 (3EK_D) *Protein ID (NP or XP #) or Wolbachia#: *Organism (including strain): Burkholderia Pseudomallei 1719b Etiologic Risk Group (see link below):Risk II *Background/Disease Information (sort of like the Intro to your Mini Research Write up): Burkholderia pseudomallei is classified as a category B bioterrorism pathogen by the US National Institute of Allergy and Infectious Diseases.This organism causes disease meliodosis, often refered to as Glanders, which is found in South-East Asia and parts of Northern Australia. Few cases of this disease have been reported each year, but the lack of research and medical facilities in the countries of incidence may be the cause for the underestimate in the records.Currently, there is no vaccine to prevent melioidosis. The mortality rate is very high. Treatment using first-line agents have ended in continual relapse.
Essentiality of this protein:
Fatty acid biosynthesis (FAS) is used to synthesize the metabolic precursors for membrane phospholipids in the cell wall. Due to the essential role that fatty acids play in bacterial cell survival and the low degree of sequence homology with the mammalian FAS-I synthase, the FAS-II pathway is thought to be an attractive antibacterial drug target. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3037155/?report=reader Complex of proteins?: Fatty Acid Synthesis II (FAS II) Druggable Target:
Crystal structure of eonyl-(acyl carrier protein) reductase from burkholderia pseudomallei 1719b PDB #3EK2
Current Inhibitors:
-Ceftazidime
-Imipenem Expression Information: It has been expressed in E.Coli BL21(DE3) http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3037155/?report=reader#!po=15.0000 Purification Method:
N-terminal His-tag on bpmFabI-2 was cleaved by treatment with thrombin.The purity of each protein was verified by 12% SDS–PAGE
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
MAHHHHHHMGFLDGKRILLTGLLSNRSIAYGIAKACKREGAELAFTYVGDRFKDRITEFAAEFGSELVFPCDVADDAQID
ALFASLKTHWDSLDGLVHSIGFAPREAIAGDFLDGLTRENFRIAHDISAYSFPALAKAALPMLSDDASLLTLSYLGAERA
IPNYNTMGLAKAALEASVRYLAVSLGAKGVRVNAISAGPIKTLAASGIKSFGKILDFVESNSPLKRNVTIEQVGNAGAFL
LSDLASGVTAEVMHVDSGFNAVVGGMAGLEE
*CDS Gene Sequence (paste as text only): *GC% Content for gene: *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): *GC% Content for gene (codon optimized):
*NCBI Gene # or RefSeq#:208435803 (3EK_D)
*Protein ID (NP or XP #) or Wolbachia#:
*Organism (including strain): Burkholderia Pseudomallei 1719b
Etiologic Risk Group (see link below):Risk II
*Background/Disease Information (sort of like the Intro to your Mini Research Write up):
Burkholderia pseudomallei is classified as a category B bioterrorism pathogen by the US National Institute of Allergy and Infectious Diseases.This organism causes disease meliodosis, often refered to as Glanders, which is found in South-East Asia and parts of Northern Australia. Few cases of this disease have been reported each year, but the lack of research and medical facilities in the countries of incidence may be the cause for the underestimate in the records.Currently, there is no vaccine to prevent melioidosis. The mortality rate is very high. Treatment using first-line agents have ended in continual relapse.
Essentiality of this protein:
Fatty acid biosynthesis (FAS) is used to synthesize the metabolic precursors for membrane phospholipids in the cell wall. Due to the essential role that fatty acids play in bacterial cell survival and the low degree of sequence homology with the mammalian FAS-I synthase, the FAS-II pathway is thought to be an attractive antibacterial drug target.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3037155/?report=reader
Complex of proteins?: Fatty Acid Synthesis II (FAS II)
Druggable Target:
*EC#: 1.3.1.9
Link to BRENDA EC# page:
**http://www.brenda-enzymes.org/php/result_flat.php4?ecno=1.3.1.9**
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): Spectrophotometric
Assay Information:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3037155/?report=reader#!po=15.0000
-- List cost and quantity of substrate reagents and supplier
Structure Available (PDB or Homology model)
Crystal structure of eonyl-(acyl carrier protein) reductase from burkholderia pseudomallei 1719b
PDB #3EK2
Current Inhibitors:
-Ceftazidime
-Imipenem
Expression Information:
It has been expressed in E.Coli BL21(DE3)
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3037155/?report=reader#!po=15.0000
Purification Method:
N-terminal His-tag on bpmFabI-2 was cleaved by treatment with thrombin.The purity of each protein was verified by 12% SDS–PAGE
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
MAHHHHHHMGFLDGKRILLTGLLSNRSIAYGIAKACKREGAELAFTYVGDRFKDRITEFAAEFGSELVFPCDVADDAQID
ALFASLKTHWDSLDGLVHSIGFAPREAIAGDFLDGLTRENFRIAHDISAYSFPALAKAALPMLSDDASLLTLSYLGAERA
IPNYNTMGLAKAALEASVRYLAVSLGAKGVRVNAISAGPIKTLAASGIKSFGKILDFVESNSPLKRNVTIEQVGNAGAFL
LSDLASGVTAEVMHVDSGFNAVVGGMAGLEE
*length of your protein in Amino Acids: 271
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 28795.8
Molar Extinction coefficient of your protein at 280 nm wavelength: 14565
http://web.expasy.org/cgi-bin/protparam/protparam
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
*CDS Gene Sequence (paste as text only):
*GC% Content for gene:
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):